Project Summary/Abstract Pancreatic ductal adenocarcinoma (PDA) is almost universally lethal, reflecting its typical late-stage diagnosis, when effective treatment options are limited. Pancreatic intra-epithelial neoplasias (PanINs) represent an early, localized developmental stage of pancreatic ductal adenocarcinoma (PDA), when curative surgical resection remains feasible. Understanding features of PDA precursor lesions like PanINs could generate new strategies for early PDA diagnosis and intervention. We have created a novel system for genetic modification and growth of primary human pancreatic cells to reconstitute the development of lesions with molecular and pathological features of human PanINs. Upon orthotopic transplantation these immortalized clones produce stable human induced pancreatic intra-epithelial neoplasias (hiPanINs). Moreover, these PanIN lesions appear to be stable: after 6 months, they do not invade locally or metastasize. Here we propose to validate and optimize this new system for generating stable human PanIN-like lesions to enhance its uses for addressing unmet translational needs. In Aim 1 we propose to generate new hiPanIN clones whose mutational spectrum matches that found in native PanIN and PDA development. We will also assess prioritized biomarkers identified in the hiPanIN system in native human PanIN and PDA specimens. In Aim 2 we propose collaborative studies to test whether pancreatitis, further genetic modification, or extrinsic signals relevant to PDA can promote invasive or metastatic PDA in the hiPanIN orthotopic transplantation model. In summary our proposal is highly responsive to the FOA (PAR 16-059). We have created a ?novel model to fill critical gaps in translational requirements? using ?human and mammalian oncology models and their genetics?, and proposed studies to ?ensure their appropriateness for addressing unmet translational needs.?